Value of Chemoresistant Testing
for Cancer Treatment
Dr. Kai Schulze-Forster
Introduction
Personalized Medicine
.... Because humans are different!
The Problem
The response to chemotherapy (solid tumors) is about
30% !
Principles and Practice of Oncology (Cancer: Principles &
Practice (DeVita, 8th Revised edition (REV).
Lippincott Williams & Wilkins, 1. Mai 2008
Biomarker
Which
Drug?
Responder
Non-Responder
severe
side effects
Biomarker
High need for predictive biomarkers!
There is progress especially for biologicals:
- Her2
- K-RAS mutation
- EGFR mutation
- …..
Chemoresistant Assay
… but for chemotherapy ??
validated biomarkers are not available
Solution: functional bioassays
Chemoresistant Assay
Well-known method in bacterial infections:
Antibiogram
Replace bacteria by tumor cells
Oncobiogram
= Chemoresistant (Chemosensitivity) Assay
Chemoresistant Assay
Method
1. Collect tumor cells (from primary tumor, metastases
or ascites during surgery, biopsy or punction)
2. Prepare single cell suspension
3. Perform proliferation assay with drugs 1-3 days
4. Determine number of living cells
5. Calculate growth inhibition
Chemoresistant Assay
96 well Microtiter plate
- 6 concentrations
- Duplicates
- Untreated control
(negative control)
1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
G
H
Drug1
Positive Control
Drug2
Negative Control
Buffer
Chemoresistant Assay
Method
Readout: ATP determination using chemoluminescence
Very sensitive: 40 cells/well
also named ATP-TCA (ATP-Tumor Chemosensitivity Assay)
ATP + Luciferin + O2
Luciferase
AMP + 2P + CO2 + Light
Chemoresistant Assay
Results: Chart
120
Tumor Growth Inhibition %
Carboplatin
Docetaxel
100
80
60
40
20
0
200
100
50
25
Test Drug Concentration %
12,5
6,25
Chemoresistant Assay
Results:
Table
Chemoresistant Assay
Advantages
1. In vitro-testing does not affect the patient
2. several drugs and combinations can be tested in
parallel
3. hyperthermia can be included
Chemoresistant Assay
Advantages
 Safe prediction of resistance
 Avoidance of ineffective therapies
 Avoidance of unnecessary side effects
Publications
Publication search in PubMed 11/2013
 Chemosensitivity Assay : 3089 entries
 Chemosensitivity Testing Cancer: 534 entries
 ATP-TCA: 60 entries
Publications
Cancer Chemotherapy and Pharmacology
May 2012, Volume 69, Issue 5, pp 1307-1314
Predicting platinum resistance in primary advanced ovarian cancer
patients with an in vitro resistance index
Thea Eline Hetland, Janne Kærn, Martina Skrede,
Berit Sandstad, Claes Tropé, Ben Davidson,Vivi Ann Flørenes
Departement of Gynecologic Oncology, Norwegian Radium Hospital, Oslo
University Hospital, 0424 Oslo, Norway. theaeline@gmail.com
Publications
PURPOSE:
We aimed to identify primary platinum resistance in epithelial ovarian cancer (OC)
patients with FIGO stage III-IV disease by an in vitro drug-response assay and to
correlate the findings with clinical response. (…)
METHODS:
We combined the ATP-based tumor-chemosensitivity and the extreme drug
resistance assays for testing of 85 biopsies from 58 patients. Tumors were classified
as sensitive or resistant by a resistance index (RI). (…) Results were analyzed for
association with clinical platinum resistance, progression-free survival (PFS), and
overall survival (OS).
Publications
RESULTS:
RI <= 250 predicted primary platinum resistance, without misclassification of
sensitive patients. The test sensitivity for primary tumors was 15/15, specificity 3/10,
negative predictive value 3/3, and positive predictive value 15/22. Patients with in
vitro platinum-resistant samples had shorter PFS compared with patients with
sensitive samples (3.4 vs. 10.0 months, p = 0.02). Comparing patient-matched
primary and metastatic samples, there was about 1/3 mismatch in resistance. RI for
platinum was lower in primary tumors exposed to neoadjuvant chemotherapy than in
chemo-naïve tumors (p < 0.01).
CONCLUSIONS:
This in vitro assay predicted primary platinum resistance, without misclassification
of sensitive OC patients, and the results were significantly associated with PFS. We
suggest that samples from primary tumor and metastatic samples have different
responses to chemotherapy and that exposure to chemotherapy might induce in vitro
platinum resistance.
Publications
Recent Results Cancer Res. 2003;161:221-30.
ATP chemosensitivity testing in ovarian and breast cancer: early
clinical trials.
Kurbacher CM, Grecu OM, Stier U, Gilster TJ, Janát MM, Untch M, Konecny G,
Bruckner HW, Cree IA.
Division of Clinical and Experimental Gynecologic Oncology, Department of
Gynecology and Obstetrics, University of Cologne Medical Center, Kerpener Strasse
34, 50931 Köln, Germany. Christian.Kurbacher@medizin.uni-koeln.de
Publications
ABSTRACT
(…) Currently, the ATP-based tumor chemosensitivity assay (ATP-TCA) can be
regarded as the most sophisticated assay to investigate both solid samples and
effusions derived from patients with various organ tumors. (…)
Clinical trials that have been set up in heavily pretreated patients with recurrent
ovarian or breast cancer have convincingly confirmed the high activity of these
combinations previously demonstrated in preclinical investigations using the ATPTCA. In a recent phase II trial performed in 59 patients with relapsed ovarian
carcinoma, ATP-TCA-directed therapy was able to triple the response rate and to
double the survival time, compared with published empirical chemotherapy regimes.
(…)
Publications
Methods Mol Med. 2005;110:101-20.
Chemosensitivity testing using microplate adenosine triphosphatebased luminescence measurements.
Kurbacher CM, Cree IA.
Department of Gynecology and Obstertrics, University of Cologne, Cologne,
Germany.
Publications
ABSTRACT
(…) Among these, the ATP-TCA has gained particular merits for ex vivo
chemosensitivity testing of native nonhematological tumors including cancers of the
breast, ovary, gastrointestinal tract, cervix and corpus uteri, and lung; malignant
melanomas; gliomas; sarcomas; and mesotheliomas. For this indication, the ATPTCA can now be considered the best documented and validated technology. (…)
In ovarian and breast carcinomas, the predictive accuracy is > 90%, with a positive
predictive value of 85-90% and a negative predictive value near 100%, respectively.
In primary ovarian cancers, the ATP-TCA has been found to accurately predict both
clinical response and survival.
Case Report
Man, 59 year old
Nasopharynx Carcinoma
April 2010 Surgery and Radiation
Mar 2011 Metastatic Prostate Ca
PSA > 500 ng/ml, Gleason 8, Start of antiandrogenic
therapy (Fugerel, LHRH)
Case Report
April 2011 Estracyt, Zometa, PSA decreased to 21,
good general condition
May-July 2011 Zometa and hormons, PSA = 9
Aug 2011 Zometa, well-being, PSA = 22
Oct 2011 PSA=296, start Second line chemotherapy
Taxotere/ Zometa/ Prednisolon
Case Report
Dec 2011 Therapy well tolerated, EPO
Jan 2012 Relapse Nasopharynx Ca
Feb 2012 Chemoresistant assay:
Cisplatin / 5-FU / Taxans found as effective drugs
Mar 2012 Start chemotherapy (Cisplatin/5-FU/Taxotere)
Aug 2012 After 6 cycles good general condition
Case Report
Sep 2012 Progression of Nasopharynx Ca
Oct 2012 Patient dead
Summary: using the chemoresistant assay a well
tolerated chemotherapy could be found for a multiple
treated patient.
About 6 month of life prolongation
Hyperthermia
To simulate a hyperthermia we included a 1 hour
treatment at 42 °C at incubation start.
In parallel the same drugs were tested without
hyperthermia.
Case Report HT1
Woman, 72 years old
GIST, rectum
Pretreated with Glivec
Hyperthermia
Hyperthermia
Hyperthermia
Case Report HT2
Woman, 34 years old
Hyperthermia
Hyperthermia
Summary
Chemoresistant assays are useful tools
to exclude non-responding
chemotherapies, especially in pretreated
multiresistant patients.
Zentrum für molekulare Onkologie GmbH
Im Biotechnologiepark  14943 Luckenwalde
Germany (near Berlin)
Thank you
for your
attention !